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| | FKBP12, GST-Fusion, Human, Recombinant, E. coli |
|  | Cat. No. 325902 |
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FK506 binding protein 1A, 12 kDa
FKBP1A Liquid. In PBS, 10% glycerol, pH 7.2. AVOID FREEZE/THAW CYCLES. Full-length recombinant, human FK506 binding protein 12-rapamycin associated protein 1 (FKBP12) (accession number = NP000792) fused to GST at the N-terminus and expressed in E.coli. FKBP12 is a cytosolic receptor for both rapamycin and FK506 that also serves as a natural cofactor for rapamycin's inhibition of mTOR. The molecular weight of 38,000 Da includes the GST moiety. Purity: ≥90% by SDS-PAGE. One vial is enough for inhibitor screening of four K-LISA™ mTOR Activity Kits (CBA055) (400 wells).Upon initial thawing, aliquot and store at -70° C. Ref.: Jin, Y.J., et al. 1991. Proc Natl Acad Sci USA 88, 6677. |
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The activity of mTOR was assessed without inhibitor and in the presence of rapamycin alone (Cat. No. 553210), FKBP12 alone (Cat. No. 325902), FKBP12/rapamycin complex, and Wortmannin (Cat. No. 681675), using the K-LISA™ mTOR Activity Kit (Cat. No. CBA055) as outlined in the Detailed Protocol. The FKBP12/rapamaycin inhibitor complex was pre-assembled by mixing 62.5 µg/ml FKBP12 and 50 µM rapamycin on ice for 10 min. The mTOR Standard (50 µl) was incubated for 15 min on ice with FKBP12 (2 µl: final concentration 2.5 µg/ml), rapamycin (2 µl; final concentration 2 µM). FKPP12/rapamycin complex (final concentration 2.5 µg/ml, 2 µM rapamycin) or wortmannin (final concentration 10 µM). The kinase reaction was initiated by transferring each mixture to a corresponding well containing 2X kinase buffer and ATP. The results were assessed as outlined in the user protocol.
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